In vitro migration and proliferation (“wound healing”) potential of mesenchymal stromal cells generated from human CD271+ bone marrow mononuclear cells

Mesenchymal stromal cells (MSCs) are a promising cell source for tissue engineering and regenerative medicine. In our lab, we found that MSC preparations from bone marrow of many different donors had a limited capacity of in vitro differentiation into osteogenic and chondrogenic lineages—a capacity claimed to be inherent to MSCs. The current study was designed to test the hypothesis that the amount of heparin used as anticoagulant during bone marrow harvest had an inhibitory influence on the in vitro differentiation capacity of isolated MSCs. Bone marrow was obtained from the femoral cavity of twelve donors during total hip arthroplasty in the absence or presence of heparin. No coagulation was observed in the absence of heparin. The number of mononuclear cells was independent of heparin addition. Isolated MSCs were characterized by morphology, population doubling times, expression of cell surface antigens and in vitro differentiation. Results of these analyses were independent of the amount of heparin. Transcriptome analyses of cells from three randomly chosen donors and quantitative realtime PCR (qRT-PCR) analysis from cells of all donors demonstrated no clear effect of heparin on the transcriptome of the cells. This excludes heparin as a potential source of disparate results.

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Cryopreserved human bone marrow mononuclear cells as a source of mesenchymal stromal cells: application in osteoporosis research

Cytotherapy ◽

10.1080/14653240802192644 ◽

2008 ◽

Vol 10 (5) ◽

pp. 460-468 ◽

Cited By ~ 33

Author(s):

A. Casado-Díaz ◽

R. Santiago-Mora ◽

R. Jiménez ◽

J. Caballero-Villarraso ◽

C. Herrera ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Mononuclear Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Bone Marrow Mononuclear Cells ◽

Osteoporosis Research

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Isolation, expansion and differentiation of mesenchymal stromal cells from rabbits' bone marrow

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2016000500012 ◽

2016 ◽

Vol 36 (5) ◽

pp. 423-430 ◽

Cited By ~ 4

Author(s):

Renato B. Eleotério ◽

Rodrigo V. Sepúlveda ◽

Emily C.C. Reis ◽

Fabrício L. Valente ◽

Andréa P.B. Borges

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Tissue Repair ◽

Mononuclear Cells ◽

Culture Media ◽

Rabbit Model ◽

Proliferation Rate ◽

Media Formulation

Abstract: Tissue engineering has been a fundamental technique in the regenerative medicine field, once it permits to build tri-dimensional tissue constructs associating undifferentiated mesenchymal cells (or mesenchymal stromal cells - MSCs) and scaffolds in vitro. Therefore, many studies have been carried out using these cells from different animal species, and rabbits are often used as animal model for in vivo tissue repair studies. However, most of the information available about MSCs harvesting and characterization is about human and murine cells, which brings some doubts to researchers who desire to work with a rabbit model in tissue repair studies based on MSCs. In this context, this study aimed to add and improve the information available in the scientific literature providing a complete technique for isolation, expansion and differentiation of MSCs from rabbits. Bone marrow mononuclear cells (BMMCs) from humerus and femur of rabbits were obtained and to evaluate their proliferation rate, three different culture media were tested, here referred as DMEM-P, DMEM´S and α-MEM. The BMMCs were also cultured in osteogenic, chondrogenic and adipogenic induction media to prove their multipotentiality. It was concluded that the techniques suggested in this study can provide a guideline to harvest and isolate MSCs from bone marrow of rabbits in enough amount to allow their expansion and, based on the laboratory experience where the study was developed, it is also suggested a culture media formulation to provide a better cell proliferation rate with multipotentiality preservation.

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Mesenchymal stromal cells derived from CD271+ bone marrow mononuclear cells exert potent allosuppressive properties

Cytotherapy ◽

10.3109/14653249.2011.605118 ◽

2011 ◽

Vol 13 (10) ◽

pp. 1193-1204 ◽

Cited By ~ 14

Author(s):

Zyrafete Kuçi ◽

Selim Kuçi ◽

Susanne Zircher ◽

Stefanie Koller ◽

Ralf Schubert ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Mononuclear Cells ◽

Bone Marrow Mononuclear Cells

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Bone Marrow Microenvironment in Light-Chain Amyloidosis: In Vitro Expansion and Characterization of Mesenchymal Stromal Cells

Biomedicines ◽

10.3390/biomedicines9111523 ◽

2021 ◽

Vol 9 (11) ◽

pp. 1523

Author(s):

Chiara Valsecchi ◽

Stefania Croce ◽

Alice Maltese ◽

Lorenza Montagna ◽

Elisa Lenta ◽

...

Keyword(s):

Bone Marrow ◽

Cell Line ◽

Plasma Cell ◽

Mesenchymal Stromal Cells ◽

Light Chain ◽

Stromal Cells ◽

Mononuclear Cells ◽

Light Chain Amyloidosis

Immunoglobulin light-chain amyloidosis (AL) is caused by misfolded light chains produced by a small B cell clone. Mesenchymal stromal cells (MSCs) have been reported to affect plasma cell behavior. We aimed to characterize bone marrow (BM)-MSCs from AL patients, considering functional aspects, such as proliferation, differentiation, and immunomodulatory capacities. MSCs were in vitro expanded from the BM of 57 AL patients and 14 healthy donors (HDs). MSC surface markers were analyzed by flow cytometry, osteogenic and adipogenic differentiation capacities were in vitro evaluated, and co-culture experiments were performed in order to investigate MSC immunomodulatory properties towards the ALMC-2 cell line and HD peripheral blood mononuclear cells (PBMCs). AL-MSCs were comparable to HD-MSCs for morphology, immune-phenotype, and differentiation capacities. AL-MSCs showed a reduced proliferation rate, entering senescence at earlier passages than HD-MSCs. The AL-MSC modulatory effect on the plasma-cell line or circulating plasma cells was comparable to that of HD-MSCs. To our knowledge, this is the first study providing a comprehensive characterization of AL-MSCs. It remains to be defined if the observed abnormalities are the consequence of or are involved in the disease pathogenesis. BM microenvironment components in AL may represent the targets for the prevention/treatment of the disease in personalized therapies.

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Phenotypic and Functional Characterization of Mesenchymal Stromal Cells Derived from CD271+- Bone Marrow Mononuclear Cells and Mesenchymal Stromal Cells Derived through Plastic Adherence.

Blood ◽

10.1182/blood.v110.11.1921.1921 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1921-1921

Author(s):

Selim Kuci ◽

Kathrin Puetsch ◽

Zyrafete Kuci ◽

Sabine Huenecke ◽

Hermann Kreyenberg ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Mononuclear Cells ◽

Functional Characterization ◽

In Vivo Studies ◽

Bone Marrow Mononuclear Cells ◽

Cell Surface Antigens ◽

Differentiation Potential ◽

Proliferation Capacity

Abstract Mesenchymal stromal cells (MSCs) are non- hematopoietic multipotent cells which can be derived from bone marrow mononuclear cells either by plastic adherence (PA-MSCs) or by a positive selection with antibodies against cell surface antigens expressed by MSC-progenitor cells (CD271, CD73, CD146, CD105, CD166, SSEA-4 and recently GD2). In this study, we compare the phenotype, proliferation potential, differentiation potential, the cytokine expression pattern and inhibitory potential of MSC- derived by plastic adherence and MSCs derived from positively selected CD271+ bone marrow mononuclear cells (BM-MNCs). According to CFU-F assay, the enriched CD271+ BM cells possess a significantly higher frequency (2952 per 1×106 BM cells) compared to PA-MSCs (21 per 1×106 BM cells). Phenotypically, both populations expressed high levels of common MSC antigens such as CD73, CD105, CD44, CD166, CD90, HLA-Class I and were negative for CD34, CD133, and CD14. Compared to PA-MSCs, CD271+ BM cells after the isolation express high levels of the hematopoietic antigen CD45 which is down- regulated within the first passage and HLA-DR, which remains constant through many passages. These cells have a 10- to 1000-fold higher proliferation capacity compared to PA- MSCs. However, both populations differentiated in vitro along adipogenic, chondrogenic and osteogenic lineage. In MLR, both populations significantly suppressed the proliferation of PHA- stimulated allogeneic T- lymphocytes at the ratio 10: 1 (MSCs:T-cells). However, CD271+ BM- derived MSCs were more efficient in secreting IFN-γ, IL-1β, IL-2, IL-4, GM-CSF and TNF-α, whereas PA- MSCs secreted significantly more IL-6 and IL-8 than CD271+ BM- derived MSCs. Ongoing in vivo studies with immunodeficient NOD/SCID mice will show the whole potential of both population in the improvement of engraftment of mobilized peripheral blood hemaatopoietic CD133+ cells. Based on their higher frequency and proliferation capacity we suggest that CD271+ BM- cells may represent a better source than PA- MSCs in order to generate bulk quantities of MSCs for clinical applications.

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Growth characteristics of human bone marrow mesenchymal stromal cells at cultivation on synthetic polyelectrolyte nanofilms in vitro

Heliyon ◽

10.1016/j.heliyon.2021.e06517 ◽

2021 ◽

Vol 7 (3) ◽

pp. e06517

Author(s):

Lyudmila M. Mezhevikina ◽

Dmitriy A. Reshetnikov ◽

Maria G. Fomkina ◽

Nurbol O. Appazov ◽

Saltanat Zh. Ibadullayeva ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Growth Characteristics ◽

Human Bone ◽

Human Bone Marrow

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Evaluation of Browning Agents on the White Adipogenesis of Bone Marrow Mesenchymal Stromal Cells: A Contribution to Fighting Obesity

Cells ◽

10.3390/cells10020403 ◽

2021 ◽

Vol 10 (2) ◽

pp. 403

Author(s):

Girolamo Di Maio ◽

Nicola Alessio ◽

Ibrahim Halil Demirsoy ◽

Gianfranco Peluso ◽

Silverio Perrotta ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

White Adipocyte ◽

Drug Candidates ◽

Fat Depots ◽

Treatment Of Obesity ◽

White Fat

Brown-like adipocytes can be induced in white fat depots by a different environmental or drug stimuli, known as “browning” or “beiging”. These brite adipocytes express thermogenin UCP1 protein and show different metabolic advantages, such as the ability to acquire a thermogenic phenotype corresponding to standard brown adipocytes that counteracts obesity. In this research, we evaluated the effects of several browning agents during white adipocyte differentiation of bone marrow-derived mesenchymal stromal cells (MSCs). Our in vitro findings identified two compounds that may warrant further in vivo investigation as possible anti-obesity drugs. We found that rosiglitazone and sildenafil are the most promising drug candidates for a browning treatment of obesity. These drugs are already available on the market for treating diabetes and erectile dysfunction, respectively. Thus, their off-label use may be contemplated, but it must be emphasized that some severe side effects are associated with use of these drugs.

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